METHOD FOR DETERMINING NUCLEIC ACID BY PCR, METHOD FOR MEASURING NUMBER OF MICROORGANISM OR CELL, NUMBER OF SPECIFIC GENE AND NUMBER OF SPECIFIC GENE COPY BY PCR AND MEASURING KIT USED FOR THESE METHODS

摘要

PURPOSE: To simply and accurately determine a nucleic acid useful for measuring the number of microorganisms by reacting a reaction solution of PCR with a coloring matter compound which is not fluorescent in liberation and is fluorescent by reaction with a duplex nucleic acid and measuring the intensity of the obtained fluorescence. ;CONSTITUTION: A nucleic acid specimen is subjected to PCR in the presence of a primer required for amplification of a specific sequence region of the target nucleic acid. In the case of the target nucleic acid existing in the specimen, an amplification product comprising the amplified duplex is reacted with a coloring matter compound of formula I [a group of formula II (X is O, S, Se or Te) is pyrylium (analog) ring; R¹ and R² are H, a halogen, a (substituted) lower alkyl, etc.; R³ is a (substituted) aryl, etc.] such as 2,4-bis(N,N-dimethylaminophenyl)-6-methylpyrylium salt which is not fluorescent in liberation and is fluorescent by reaction with a duplex nucleic acid to give fluorescence. The intensity of the fluorescence is measured and the amount of the target nucleic acid in the nucleic acid specimen is obtained to determine the nucleic acid.;COPYRIGHT: (C)1995,JPO